Methods and systems for delivery of zw1 heptapeptide

ABSTRACT

The present invention generally relates to systems and methods for the transdermal delivery of the heptapeptide ZW1. In some aspects, ZW1 may be contained with a composition comprising a lecithin, such as phosphatidylcholine. In certain embodiments, the lecithin is present in liposomes, micelles, liquid crystals, or other configurations. The composition can take the form of a gel, a cream, a lotion, an ointment, a solution, a solid “stick,” etc., that can be rubbed or sprayed onto the skin. Other aspects of the present invention are generally directed to methods of making or using such compositions, methods of promoting such compositions, kits including such compositions, or the like.

FIELD

The present invention generally relates to systems and methods for thetransdermal delivery of the heptapeptide ZW1.

BACKGROUND

ZW1 is a recently discovered heptapeptide having the structure SMSARQL(SEQ ID NO: 1). ZW1 may be useful for the treatment of certainconditions such as Alzheimer's disease. ZW1 may also be useful forreducing the inflammatory response, or for improving memory, e.g.,spatial memory. However, ZW1 is highly charged, and is usually given viadirect injection into the brain. Accordingly, improvements in thedelivery of ZW1 are needed.

SUMMARY

The present invention generally relates to systems and methods for thetransdermal delivery of the heptapeptide ZW1. The subject matter of thepresent invention involves, in some cases, interrelated products,alternative solutions to a particular problem, and/or a plurality ofdifferent uses of one or more systems and/or articles.

In one aspect, the present invention is generally directed to an articlefor transdermal delivery. In one set of embodiments, the articlecomprises a composition comprising a carrier and lecithin, the lecithincomprising ZW1 (SEQ ID NO: 1). In some cases, the composition furthercomprises no more than about 250 ppm of water by weight of thecomposition.

The article, in another set of embodiments, comprises a compositioncomprising a carrier and lecithin, the lecithin comprising ZW1 (SEQ IDNO: 1). In some cases, the composition is stable at room temperature.

The present invention, in another aspect, is generally directed to amethod. According to one set of embodiments, the method comprisesadministering, to a subject, a composition comprising a carrier andlecithin, the lecithin comprising ZW1 (SEQ ID NO: 1). In someembodiments, the composition further comprises no more than about 250ppm of water by weight of the composition.

The method, in another set of embodiments, includes administering, to asubject, a composition comprising a carrier and lecithin, the lecithincomprising ZW1 (SEQ ID NO: 1). In some cases, the composition is stableat room temperature.

Several methods are disclosed herein of administering a subject with acompound for prevention or treatment of a particular condition. It is tobe understood that in each such aspect of the invention, the inventionspecifically includes, also, the compound for use in the treatment orprevention of that particular condition, as well as use of the compoundfor the manufacture of a medicament for the treatment or prevention ofthat particular condition.

In another aspect, the present invention encompasses methods of makingone or more of the embodiments described herein, for example, acomposition comprising ZW1. In still another aspect, the presentinvention encompasses methods of using one or more of the embodimentsdescribed herein, for example, a composition comprising ZW1.

Other advantages and novel features of the present invention will becomeapparent from the following detailed description of various non-limitingembodiments of the invention. In cases where the present specificationand a document incorporated by reference include conflicting and/orinconsistent disclosure, the present specification shall control. If twoor more documents incorporated by reference include conflicting and/orinconsistent disclosure with respect to each other, then the documenthaving the later effective date shall control.

BRIEF DESCRIPTION OF THE SEQUENCES

SEQ ID NO: 1 is SMSARQL or ZW1.

DETAILED DESCRIPTION

The present invention generally relates to systems and methods for thetransdermal delivery of the heptapeptide ZW1. In some aspects, ZW1 maybe contained with a composition comprising a lecithin, such asphosphatidylcholine. In certain embodiments, the lecithin is present inliposomes, micelles, liquid crystals, or other configurations. Thecomposition can take the form of a gel, a cream, a lotion, an ointment,a solution, a solid “stick,” etc., that can be rubbed or sprayed ontothe skin. Other aspects of the present invention are generally directedto methods of making or using such compositions, methods of promotingsuch compositions, kits including such compositions, or the like.

In certain aspects, the present invention is generally directed tocompositions comprising ZW1 for application to the skin of a subject,e.g., a human subject. In some embodiments, the composition is generallydirected to a cream, or other formulation, that contains a lecithin,such as phosphatidylcholine. The composition may be applied to the skinof a subject, such as a human subject, to treat a variety of diseases orconditions. For example, in one set of embodiments, the composition maybe used to treat a subject having Alzheimer's disease. Other examples oftreatment of various diseases or conditions are discussed in additionaldetail below.

In one set of embodiments, compositions such as those described hereinmay be used to deliver ZW1 across the skin into the bloodstream. Oncedelivered into the bloodstream, ZW1 may inherently be able to cross theblood-brain barrier, e.g., due to its small size. Delivering ZW1 throughthe skin avoids degradation problems in oral delivery, and thedifficulties in directly delivering ZW1 into the brain.

ZW1 may be prepared by any suitable technique, including synthetictechniques such as solid-phase synthesis techniques, liquid-phasesynthesis techniques, etc. ZW1 may be prepared by recombinant methods insome embodiments.

Any suitable amount of ZW1 may be present within a composition preparedas described herein. For example, at least about 0.3 wt %, at leastabout 0.5 wt %, at least about 0.7 wt %, at least about 1 wt %, at leastabout 1.5 wt %, at least about 2 wt %, at least about 2.5 wt %, at leastabout 3 wt %, at least about 5 wt % at least about 10 wt %, at leastabout 20 wt %, at least about 30 wt %, at least about 40 wt %, at leastabout 50 wt %, at least about 60 wt %, at least about 70 wt %, at leastabout 80 wt %, at least about 90 wt %, at least about 100 wt %, at leastabout 110 wt %, or at least about 120 wt % of the composition can beZW1, where the basis of the weight percentage is the weight of thecomposition before the ZW1 is added. Combinations of any of these arealso possible. For example, ZW1 may be present at between 70 wt % andabout 120 wt % of the composition.

In some embodiments, ZW1 may be present at a concentration of at leastabout 400 mg/kg, at least about 450 mg/kg, at least about 500 mg/kg, atleast about 550 mg/kg, at least about 570 mg/kg, at least about 600mg/kg, at least about 650 mg/kg, at least about 700 mg/kg, at leastabout 750 mg/kg, at least about 800 mg/kg, at least about 850 mg/kg, atleast about 950 mg/kg, or at least about 1000 mg/kg of the composition,where the mass that is determined is that of the final composition,e.g., including ZW1. In certain cases, the ZW1 may be present at aconcentration of no more than about 2000 mg/kg, no more than about 1500mg/kg, no more than about 1000 mg/kg, no more than about 960 mg/kg, nomore than about 900 mg/kg, no more than about 800 mg/kg, no more thanabout 700 mg/kg, or no more than about 600 mg/kg. Combinations of any ofthese are also possible. For example, the ZW1 may be present at aconcentration of between about 570 mg/kg and about 960 mg/kg.

In some embodiments, ZW1 is present at a concentration (e.g., on aper-mass basis) of at least about 100 ppm, at least about 200 ppm, atleast about 300 ppm, at least about 400 ppm, at least about 500 ppm, atleast about 600 ppm, at least about 700 ppm, at least about 800 ppm, atleast about 900 ppm, at least about 1000 ppm, at least about 1100 ppm,at least about 1200 ppm, at least about 1300 ppm, at least about 1400ppm, at least about 1500 ppm, at least about 1600 ppm, at least about1700 ppm, at least about 1800 ppm, at least about 1900 ppm, at leastabout 2000 ppm, at least about 2500 ppm, at least about 3000 ppm, atleast about 3500 ppm, at least about 4000 ppm, at least about 4500 ppm,at least about 5000 ppm, at least about 6000 ppm, at least about 7000ppm, at least about 8000 ppm, at least about 9000 ppm, or at least about10000 ppm of the composition. In other embodiments, the ZW1 is presentat a concentration of no more than about 11000 ppm, no more than about10000 ppm, no more than about 9000 ppm, no more than about 8000 ppm, nomore than about 7000 ppm, no more than about 6000 ppm, no more thanabout 5000 ppm, no more than about 4500 ppm, no more than about 4000ppm, no more than about 3500 ppm, no more than about 3000 ppm, no morethan about 2500 ppm, no more than about 2000 ppm, no more than about1900 ppm, no more than about 1800 ppm, no more than about 1700 ppm, nomore than about 1600 ppm, no more than about 1500 ppm, no more thanabout 1400 ppm, no more than about 1300 ppm, no more than about 1200ppm, no more than about 1100 ppm, no more than about 1000 ppm, no morethan about 900 ppm, no more than about 800 ppm, no more than about 700ppm, no more than about 600 ppm, no more than about 500 ppm, no morethan about 400 ppm, or no more than about 300 ppm of the composition.Combinations of any of these are also possible. For example, in someembodiments, ZW1 is present at a concentration of between about 400 andabout 900 ppm. In addition, in some cases, the composition may includeZW1 and nitric oxide (NO). The NO may be present in gaseous or molecularform.

In certain aspects of the invention, ZW1 may be contained within acomposition that is to be applied to the skin, e.g., such that ZW1 maybe administered for topical or transdermal delivery. In some cases, thecomposition is a cream, although other formulations are also possible insome instances, e.g., a liquid, a gel, a lotion, an ointment, a solid“stick,” or the like, such as is discussed herein.

In certain embodiments of the present invention, ZW1 may be containedwithin certain compositions comprising lecithin or phosphatidylcholine.In one set of embodiments, the composition may comprise liquid crystalmultilamellar phosphatidylcholine. In some cases, the compositions maybe stable, and/or can be stored for periods of time with little or noloss or reaction of the ZW1 contained therein. In some cases, stabilityof the composition can be achieved at room temperature (about 25° C.),and/or at other storage temperatures such as those described herein.

In some embodiments, ZW1 may be added to a composition during or afterthe formulation of any phase or composition as described herein, e.g.,by routine methods known in the art. ZW1 is highly charged and generallybelieved to be hydrophilic. For example, ZW1 may be added to any phaseof a formulation or composition, or after any formulation or compositiondescribed herein is made. In some cases, for example, ZW1 may be addedbefore or after nitric oxide is added to the mixture, or before or afterthe first and second phases are mixed together. ZW1 may be present in ahydrophilic portion (e.g., a first portion) of a formulation. Forexample, if the formulation is an emulsion, ZW1 may be contained withina hydrophilic phase of the emulsion.

In one set of embodiments, the composition comprises a first phasecomprising a lecithin such as phosphatidylcholine, which may be presentwithin a second phase comprising an emulsifier, such as is discussedherein. Other components, for example, transdermal penetrationenhancers, adjuvants, surfactants, lubricants, etc. can also be presentin certain cases.

The compositions of the invention comprise, in certain embodiments, aphase comprising phosphatidylcholine and/or other lecithins in which ZW1may be contained within. In some cases, the phosphatidylcholine orlecithin may be contained within a second phase, for example, comprisingan emulsifier, which may cause the phosphatidylcholine or lecithin toform vesicles, e.g., micelles or liposomes. The phosphatidylcholine orlecithin composition can be unilamellar or multilamellar in someembodiments. However, in some instances, the presence of the secondphase causes the phosphatidylcholine or lecithin to form a liquidcrystal arrangement, rather than a vesicular or liposomal arrangement.

Without wishing to be bound by any theory, it is believed that thephosphatidylcholine or other lecithin may be used to surround the ZW1.In some cases, the ZW1 may be contained within water or other aqueousenvironment within the composition (e.g., within vesicles such asliposomes or an emulsion within the composition, etc.), although in someembodiments, little or no water is used and the ZW1 is directlycontained within the phosphatidylcholine or other lecithin within thecomposition.

For example, in certain embodiments of the invention, the composition,or at least a phase of the composition comprising ZW1 is substantiallyfree of water, e.g., comprising no more than about 10 wt %, no more thanabout 3 wt %, no more than about 1 wt %, no more than about 0.3 wt %, orno more than about 0.1 wt % water (i.e., relative to the weight of theoverall composition). The composition may also have no more than about1,000 ppm, no more than about 750 ppm, no more than about 500 ppm, nomore than about 400 ppm, no more than about 300 ppm, no more than about250 ppm, no more than about 200 ppm, no more than about 150 ppm, no morethan about 100 ppm, no more than about 50 ppm, no more than about 25ppm, or no more than about 10 ppm of water (by weight). In certainembodiments, no detectable water may be present in the composition, orat least within a phase of the composition comprising the ZW1. Anysuitable technique can be used for determining the amount of waterpresent in the composition, for example, Karl-Fisher titration. In somecases, the composition may also be free of any liquids that typicallycontain water, e.g., physiological buffers, bodily fluids, saline,blood, or the like.

In addition, in some embodiments, the composition, or at least a phaseof the composition comprising ZW1, etc. is substantially free of gaseousoxygen (O₂). For instance, the composition may also have no more thanabout 1,000 ppm, no more than about 750 ppm, no more than about 500 ppm,no more than about 400 ppm, no more than about 300 ppm, no more thanabout 250 ppm, no more than about 200 ppm, no more than about 150 ppm,no more than about 100 ppm, no more than about 50 ppm, no more thanabout 25 ppm, or no more than about 10 ppm of oxygen (by weight).

Phosphatidylcholine (herein abbreviated “PC”) is a basic component ofcell membrane bilayers and the main phospholipid circulating in theplasma of blood. Phosphatidylcholine typically has a phospholipidstructure with a choline head group and a glycerophosphoric acid tailgroup. The tail group can be saturated or unsaturated. More than onetail group may be present in the phosphatidylcholine in some cases, andthe tail groups may be the same or different. Specific non-limitingexamples of phosphatidylcholines that could be used include one or amixture of stearic, palmitic, margaric, and/or oleic acid diglycerideslinked to a choline ester head group.

Phosphatidylcholines are a member of a class of compounds calledlecithins. Typically, a lecithin is a composed of phosphoric acid,choline, fatty acids, glycerol, glycolipids, triglycerides, and/orphospholipids. In some cases, other lecithins may be used, in additionto or instead of a phosphatidylcholine. Non-limiting examples of otherlecithins include phosphatidylethanolamine, phosphatidylinositol, orphosphatidic acid. Many commercial lecithin products are available, suchas, for example, Lecithol®, Vitellin®, Kelecin®, and Granulestin®.Lecithin is widely used in the food industry. In some embodiments,certain compositions of the invention can contain synthetic or naturallecithin, or mixtures thereof. Natural preparations are used in somecases because they exhibit desirable physical characteristics, and/ormay be economical or nontoxic. However, in other embodiments,non-natural preparations are used, or the composition can include bothnatural and non-natural preparations.

Any suitable amount of phosphatidylcholine or lecithin may be presentwithin the composition. For example, at least about 0.25 wt %, at leastabout 0.5 wt %, at least about 1 wt %, at least about 2 wt %, at leastabout 3 wt %, at least about 5 wt %, at least about 8 wt %, at leastabout 10 wt %, at least about 20 wt %, at least about 30 wt %, at leastabout 40 wt %, at least about 50 wt %, at least about 60 wt %, at leastabout 70 wt %, at least about 80 wt %, or at least about 90 wt % of theentire composition can be a phosphatidylcholine or a lecithin. In somecases, the phosphatidylcholine or lecithin may be present at aconcentration of no more than about 95 wt %, no more than about 90 wt %,no more than about 80 wt %, no more than about 70 wt %, no more thanabout 65 wt %, no more than about 60 wt %, no more than about 50 wt %,no more than about 40 wt %, no more than about 30 wt %, no more thanabout 20 wt %, or no more than about 10%. Combinations of any of theseare also possible. For instance, the phosphatidylcholine or lecithin maybe present at between about 8 wt % and about 65 wt %, or between about 0wt % and about 10 wt %, etc. One or more than one type ofphosphatidylcholine or lecithin may be present.

In some aspects, the formulation comprises a phosphatidylcholine, e.g.,any of those described herein. The composition can include any suitableamount of phosphatidylcholine, for example, at least about 1 wt %, atleast about 3 wt %, at least about 5 wt %, at least about 10 wt %, atleast about 20 wt %, at least about 30 wt %, at least about 40 wt %, atleast about 50 wt %, at least about 60 wt %, at least about 70 wt %, atleast about 80 wt %, at least about 90 wt % etc. In some cases, no morethan about 90 wt %, no more than about 80 wt %, no more than about 70 wt%, no more than about 60 wt %, no more than about 50 wt %, no more thanabout 40 wt %, no more than about 30 wt %, no more than about 20 wt %,no more than about 10 wt %, or no more than about 5 wt % of thecomposition is phosphatidylcholine. Combinations of any of these arealso possible. For example, the composition may be between about 0 wt %and about 10 wt % surfactant. The composition may include one or morethan one phosphatidylcholine. One non-limiting example of aphosphatidylcholine is Phospholipon-90G (American Lecithin Company).

Some compositions may contain polyenylphosphatidylcholine (hereinabbreviated “PPC”). In some cases, PPC can be used to enhance epidermalpenetration. The term “polyenylphosphatidylcholine,” as used herein,means any phosphatidylcholine bearing two fatty acid moieties, whereinat least one of the two fatty acids is an unsaturated fatty acid with atleast two double bonds in its structure, such as linoleic acid.

Certain types of soybean lecithin and soybean fractions, for example,can contain higher levels of polyenylphosphatidylcholine, withdilinoleoylphosphatidylcholine (18:2-18:2 phosphatidylcholine) as themost abundant phosphatidylcholine species therein, than conventionalfood grade lecithin. Such lecithins may be useful in formulating certaindelivery compositions. In some embodiments, conventional soybeanlecithin may be enriched with polyenylphosphatidylcholine, for instance,by adding soybean extracts containing high levels ofpolyenylphosphatidylcholine. As used herein, this type ofphosphatidylcholine is called “polyenylphosphatidylcholine-enriched”phosphatidylcholine (hereinafter referred to as PPC-enrichedphosphatidylcholine), even where the term encompasses lecithin obtainedfrom natural sources exhibiting polyenylphosphatidylcholine levelshigher than ordinary soybean varieties. These products are commerciallyavailable, for example, from American Lecithin Company, Rhone-Poulencand other lecithin vendors. American Lecithin Company markets itsproducts with a “U” designation, indicating high levels of unsaturation;Rhone-Poulenc's product is a soybean extract containing about 42%dilinoleoylphosphatidylcholine and about 24%palmitoyllinoleylphosphatidylcholine (16:0 to 18:2 of PC) as the majorphosphatidylcholine components. Another example of a suitablepolyenylphosphatidylcholine is NAT 8729 (also commercially availablefrom vendors such as Rhone-Poulenc and American Lecithin Company).

Any suitable amount of polyenylphosphatidylcholine may be present withinthe composition. For example, at least about 0.25 wt %, at least about0.5 wt %, at least about 1 wt %, at least about 2 wt %, at least about 3wt %, at least about 5 wt %, at least about 8 wt %, at least about 10 wt%, at least about 20 wt %, at least about 30 wt %, at least about 40 wt%, at least about 50 wt %, at least about 60 wt %, at least about 70 wt%, at least about 80 wt %, or at least about 90 wt % of the compositioncan be polyenylphosphatidylcholine. In some cases, thepolyenylphosphatidylcholine may be present at a concentration of no morethan about 95 wt %, no more than about 90 wt %, no more than about 80 wt%, no more than about 70 wt %, no more than about 65 wt %, no more thanabout 60 wt %, no more than about 50 wt %, no more than about 40 wt %,no more than about 30 wt %, no more than about 20 wt %, or no more thanabout 10%. Combinations of any of these are also possible. For instance,the polyenylphosphatidylcholine may be present at between about 8 wt %and about 65 wt %. In some embodiments, at least about 20 wt %, at leastabout 30 wt %, at least about 40 wt %, at least about 50 wt %, at leastabout 60 wt %, at least about 70 wt %, at least about 80 wt %, at leastabout 90 wt %, or about 100 wt % of all of the phosphatidylcholine orlecithin in the composition is polyenylphosphatidylcholine.

In certain embodiments, a composition such as those described herein canbe formulated to include a first phase and a second phase. Typically,the second phase is substantially immiscible with the first phasecomprising phosphatidylcholine or lecithin. Two phases that aresubstantially immiscible are able to form discrete phases when exposedto each other at ambient conditions (e.g., 25° C. and 1 atm) forextended periods of time (e.g., at least about a day). The phases can beseparate identifiable phases (e.g., one may float above the other), orin some cases, the phases are intermingled, e.g., as in an emulsion. Thestability of the discrete phases may be kinetic and/or thermodynamic innature, in various embodiments.

In one set of embodiments, the second phase may comprise an emulsifierwhich causes the first phase comprising phosphatidylcholine or lecithinto form a liquid crystal, and/or vesicles such as micelles or liposomes.Typically, in a liquid crystal phase, vesicular structures such asmicelles, liposomes, hexagonal phases, or lipid bilayers can be formed.In some cases, multilamellar structures may be present within the liquidcrystal phase, although in other cases, only unilamellar structures maybe present. For example, in certain cases, the PPC-enrichedphosphatidylcholine can be loosely arranged in a multilamellar fashion.In some cases, the first phase (e.g., comprising PPC-enrichedphosphatidylcholine) and the second phase can form a structure such asis disclosed in U.S. Pat. No. 7,182,956 to Perricone, et al. This isbelieved (without wishing to be bound by any theory) to form a looselyarranged, yet stable, PPC-enriched phosphatidylcholine-drug complex thatmay allow penetration and delivery of ZW1 and optional adjunctingredients to the skin, e.g., to the dermal vasculature, or to amucosal surface.

Thus, in some embodiments, the second phase may comprise an emulsifier.The emulsifier, in one embodiment, may be a substance that is able tostabilize an emulsion by increasing its kinetic stability. Theemulsifier may also be chosen in some cases to be relatively inert ornon-toxic relative to the skin or to a mucosal surface.

A variety of emulsifiers can be used, and many emulsifiers are readilyavailable commercially. In one embodiment, for example, the emulsifiercomprises a surfactant. Non-limiting examples of surfactants include asiloxylated polyether comprising dimethyl, methyl(propylpolyethyleneoxide propylene oxide, acetate) siloxane commercially available fromvendors such as Dow Corning (Dow Corning 190 surfactant). Other examplesof materials that can be used as (or within) the second phase (e.g., asemulsifiers) include, but are not limited to, 1,2-propanediol, orsilicone fluids containing low viscosity polydimethylsiloxane polymers,methylparaben (p-hydroxy benzoic acid methyl ester) commerciallyavailable from vendors such as Dow Corning (Dow Corning 200 siliconefluid). Still other examples include various siloxane or siliconecompounds, e.g., hexamethyldisiloxane, amodimethicone,phenyltrimethicone, etc.

In some embodiments, the second phase may comprise a polyglycol. Thepolyglycol may include a polyhydric alcohol of a monomeric glycol suchas polyethylene glycol (PEG) and/or polypropylene glycol (PPG). Forexample, the PEG or PPG may be PEG or PPG 200, 300, 400, 600, 1,000,1,450, 3,350, 4,000, 6,000, 8,000, and 20,000, where the numberindicates the approximate average molecular weight of the PEG or PPG. Asis understood by those of ordinary skill in the art, a polyglycolcomposition often will comprise a range of molecular weights, althoughthe approximate average molecular weight is used to identify the typepolyglycol. More than one PEG and/or PPG can also be present in certaininstances.

More than one PEG and/or PPG can also be present in certain instances.The composition can include any suitable amount of polyglycol, forexample, at least about 1 wt %, at least about 3 wt %, at least about 5wt %, at least about 10 wt %, at least about 20 wt %, at least about 30wt %, at least about 40 wt %, at least about 50 wt %, etc. In somecases, no more than about 60 wt %, no more than about 50 wt %, no morethan about 40 wt %, no more than about 30 wt %, no more than about 20 wt%, no more than about 18 wt %, no more than about 15 wt %, no more thanabout 12 wt %, or no more than about 10 wt % of the composition ispolyglycol. Combinations of any of these are also possible. For example,the composition may be between about 0 wt % and about 10 wt %polyglycol. The composition may include one or more than one type ofpolyglycol.

Additionally, purified water may be added to the second phase in someembodiments, although in other cases, little or no water is present inthe second phase. For example, the first phase, the second phase, cancontain less than 10%, less than 5%, less than 2%, less than 1%, or lessthat 0.05% (e.g., wt %) of water relative to the weight of therespective phase or of the entire composition. In some cases, the secondphase may also comprise adjunct ingredients such as those describedherein.

The second phase may include any one, or more than one, of the materialsdescribed above. In addition, any suitable amount of second phase can beused in accordance with various embodiments of the invention. Forexample, the second phase may be present at at least about 10 wt %, atleast about 20 wt %, at least about 30 wt %, at least about 40 wt %, atleast about 50 wt %, at least about 60 wt %, at least about 70 wt %, atleast about 80 wt %, or at least about 90 wt % of the composition. Insome cases, the ratio of the first phase (e.g., comprisingphosphatidylcholine or lecithin) to the second phase can be at leastabout 1:3, at least about 1:2, at least about 1:1, at least about 2:1,at least about 3:1, or at least about 4:1, etc.

As a specific non-limiting example of one set of embodiments, apolyenylphosphatidylcholine comprises a certain material with the tradename NAT 8729, and optionally at least one polyglycol (e.g., PEG or PPG,such as is described herein). The composition can also comprise aPPC-enriched phosphatidylcholine material that is present within thefirst or second phase, e.g., comprising ZW1. The second phase may alsocomprise a surfactant such as a siloxylated polyether comprisingdimethyl, methyl(propylpolyethylene oxide propylene oxide, acetate)siloxane commercially available from vendors such as Dow Corning (DowCorning 190 surfactant) and lubricant such as silicone fluids containinglow viscosity polydimethylsiloxane polymers, methylparaben (p-hydroxybenzoic acid methyl ester) commercially available from vendors such asDown Corning (Dow Corning 200 silicone fluid).

Other examples of materials that can be used as (or within) theformulation include, but are not limited to, benzyl alcohol, ethylalcohol, isopropyl palmitate (IPP), propanediol, and caprylic/caprictriglycerides.

As another example, the first phase also comprises, in some embodimentsof the invention, a fatty acid ester. Non-limiting examples includeascorbate palmitate or isopropyl palmitate. In some cases, the fattyacid ester is used as a preservative or an antioxidant. The compositioncan include any suitable amount of fatty acid ester, for example, atleast about 1 wt %, at least about 3 wt %, at least about 5 wt %, atleast about 10 wt %, at least about 20 wt %, at least about 30 wt %, atleast about 40 wt %, at least about 50 wt %, etc. In some cases, no morethan about 60 wt %, no more than about 50 wt %, no more than about 40 wt%, no more than about 30 wt %, no more than about 20 wt %, no more thanabout 18 wt %, no more than about 15 wt %, no more than about 12 wt %,or no more than about 10 wt % of the composition is fatty acid ester.Combinations of any of these are also possible. For example, thecomposition may be between about 0 wt % and about 10 wt % fatty acidester. The composition may include one or more than one fatty acidester.

In another set of embodiments, the composition may also include one ormore transdermal penetration enhancers. Examples of transdermalpenetration enhancers include, but are not limited to,1,3-dimethyl-2-imidazolidinone or 1,2-propanediol. Other examplesinclude cationic, anionic, or nonionic surfactants (e.g., sodium dodecylsulfate, polyoxamers, etc.); fatty acids and alcohols (e.g., ethanol,oleic acid, lauric acid, liposomes, etc.); anticholinergic agents (e.g.,benzilonium bromide, oxyphenonium bromide); alkanones (e.g., n-heptane);amides (e.g., urea, N,N-dimethyl-m-toluamide); organic acids (e.g.,citric acid); sulfoxides (e.g., dimethylsulfoxide); terpenes (e.g.,cyclohexene); ureas; sugars; carbohydrates or other agents. Thetransdermal penetration enhancers can be present in any suitable amountwithin the composition. For example, at least about 10 wt %, at leastabout 20 wt %, at least about 30 wt %, at least about 40 wt %, or atleast about 50 wt % of the composition may comprise one or moretransdermal penetration enhancers. In some cases, no more than about 60wt %, no more than about 50 wt %, no more than about 40 wt %, no morethan about 30 wt %, no more than about 20 wt %, no more than about 10 wt%, no more than about 9 wt %, or no more than about 5 wt % of thecomposition comprises transdermal penetration enhancers. Combinations ofany of these are also possible. For example, the composition may havebetween about 0 wt % and about 5 wt % of one or more transdermalpenetration enhancers.

In other embodiments, the composition may be modified in order tocontrol depth of penetration. For example, in certain embodiments, thecomposition includes one or more polymers that act to reduce penetrationdepth of ZW1. Controlled depth of penetration may be important forindications where local administration is desired without systemiceffects. Examples of transdermal penetration barrier polymers include,but are not limited to, silicone waxes, acrylate polymers, anddimethicone copolymers. In certain embodiments, a transdermalpenetration barrier polymer is nonionic. A transdermal penetrationbarrier polymer can be present in any suitable amount within thecomposition. For example, at least about 10 wt %, at least about 20 wt%, at least about 30 wt %, at least about 40 wt %, or at least about 50wt % of the composition may comprise one or more transdermal penetrationbarrier polymers. In some cases, no more than about 60 wt %, no morethan about 50 wt %, no more than about 40 wt %, no more than about 30 wt%, no more than about 20 wt %, no more than about 10 wt %, no more thanabout 9 wt %, or no more than about 5 wt % of the composition comprisesa transdermal penetration barrier polymer. Combinations of any of theseare also possible. For example, the composition may have between about 0wt % and about 5 wt % of one or more transdermal penetration barrierpolymers.

In some embodiments, various compositions of the invention areformulated to be substantially clear or substantially transparent.Transparency may be useful, for instance, for product acceptance in themarketplace, e.g., when applied to the skin of a subject. However, inother embodiments, the composition is not necessarily transparent.Certain substances can be useful in providing a substantiallytransparent composition, for example, fatty acid esters such asascorbate palmitate or isopropyl palmitate. In one set of embodiments,the composition may be substantially transparent such that incidentvisible light (e.g., have wavelengths of between about 400 nm and about700 nm) can be transmitted through 1 cm of the composition with a lossin intensity of no more than about 50%, about 60%, about 70%, about 80%,or about 90% relative to the incident light. In some embodiments, theremay be no substantial difference in the wavelengths that are absorbed bythe composition (i.e., white light passing through the compositionappears white), although in other cases, there can be more absorption atvarious wavelengths (for example, such that white light passing throughthe composition may appear colored).

Other components may also be present within the composition, inaccordance with certain embodiments of the invention. For example, thecomposition may include volatile organic fluids, fatty acids, volatilearomatic cyclic compounds, high molecular weight hydrocarbons, or thelike.

As mentioned above, some aspects of the invention include nitric oxide(NO) in addition to ZW1. Without wishing to be bound by theory, it isbelieved that nitric oxide forms reversible physical bonds, similar tohydrogen bonds or van der Waals forces, with phosphatidylcholine orother lecithin molecules, e.g., containing one or more double bonds,which may allow nitric oxide to become entrapped and thereby remainintact for an extended period of time, e.g., during storage. Thesephysical bonds, however, are believed to be not very stable, and may insome cases be easily broken up, for example, upon various physicalagitations such as rubbing the composition against skin or a mucosalsurface, thereby releasing the entrapped nitric oxide. While others havestabilized other substances or drugs within phosphatidylcholine orlecithin compositions or vesicles, for example, protein drugs such asinsulin, it is surprising that a small, highly reactive molecule such asNO could similarly be stabilized, especially when it would have beenexpected that a molecule as small as NO would readily diffuse away fromsuch compositions and/or would have reacted with water that is typicallypresent within such compositions.

In some embodiments, nitric oxide not only can be entrapped inphosphatidylcholine or lecithin compositions such as those describedherein, but also that such entrapped compositions may have a long shelflife, especially when refrigerated. No or little loss or reaction ofnitric oxide is expected during extended refrigerated storage, at leastunder certain conditions.

Without wishing to be bound by any theory, it is believed that when someof the compositions described herein are applied to the skin, the liquidcrystal structure collapses, delivering ZW1. The concentration of ZW1inside the liquid crystal matrix can be varied in terms ofconcentration. The matrix also may act as a sustained release deliverysystem in some embodiments. It is also believed that the liquid crystalis highly penetrating, such that ZW1 can be delivered to the epidermis,dermis and dermal vascular for systemic release as well as tosubcutaneous fat, at least under some conditions.

In one set of embodiments, a composition such as is discussed herein maybe prepared and/or stored at any suitable temperature and under anysuitable conditions. In some embodiments, for instance, a compositioncan be prepared and/or stored under limited or no oxygen conditions. Thecomposition can also be prepared and/or stored under limited or nonitrogen and/or carbon dioxide. For instance, the composition may beprepared and/or stored in a sealed environment (e.g., stored in a sealedcontainer). The sealed environment (e.g., container) can be at leastsubstantially devoid of gas, and/or contains a gaseous mixture thatexcludes, or at least is depleted in, oxygen. In some embodiments, anenvironment depleted in oxygen may have less than about 20%, less thanabout 15%, less than about 10%, less than about 5%, about 1% or less,about 0.1% or less, about 0.01% or less, about 0.001% or less, oxygen(e.g., as a wt % or as molar % per volume). For example, the gaseousmixture may include a noble gas, such as argon, helium, neon, etc. Inone set of embodiments, the container may comprise a multi-layeredmetallic and/or polymeric barrier, e.g., formed from Glaminate®(American Can Company). For instance, the container may have the shapeof a tube. Thus, in certain embodiments, the container is substantiallyresistant to oxygen permeation, nitrogen permeation, and/or carbondioxide permeation. In certain embodiments, the container issubstantially watertight, for example, such that substantially no wateris absorbed by the container, or such that no water is able to passthrough the container even if the container is filled with water.

In certain embodiments, the composition may be stored at temperatures ofless than about 80° C., less than about 70° C., less than about 60° C.,less than about 50° C., less than about 40° C., less than about 30° C.,less than about 25° C., less than about 20° C., less than about 15° C.,less than about 10° C., less than about 5° C., less than about 0° C.,etc., for extended periods of time, e.g., at least about a day, at leastabout a week, at least about 4 weeks, at least about 6 months, etc.

In accordance with certain aspects of the invention, a composition asdiscussed herein may be prepared by mixing a first phase and a secondphase together. The second phase can comprise an emulsifier, or anyother components discussed herein. The first phase may comprise alecithin such as phosphatidylcholine and/or polyenylphosphatidylcholine,e.g., PPC-enriched phosphatidylcholine, for instance, as describedherein. In some embodiments, other components are also mixed into thecomposition, for example, transdermal penetration enhancers, adjuvants,polyglycols (e.g., PEG and/or PPG), surfactants, lubricants, etc. asdiscussed herein.

In some embodiments, ZW1 may be added to a composition during or afterthe formulation of any phase or composition as described herein, e.g.,by routine methods known in the art. For example, ZW1 may be added toany phase of a formulation or composition, or after any formulation orcomposition described herein is made.

In addition, in some embodiments, the composition may include nitricoxide as well. The nitric oxide may be added, for example, by passingbubbles of nitric oxide through the composition. See, for example, U.S.Pat. No. 8,668,937, incorporated herein by reference in its entirety.

In one aspect, application of ZW1 in a composition as described herein,may be applied to the skin of a subject, e.g., one having or at risk ofa disease, condition, or need described herein. Additionally, in someembodiments, the composition may be applied in conjunction with othertypes of treatments to a subject, e.g., to the skin of a subject, fortreatment of any of the diseases, conditions, or needs described herein.These may be occur, e.g., simultaneously or sequentially, in variousembodiments. Thus, certain compositions as described herein may be usedto treat a wide variety of diseases or conditions. To “treat” a disordermeans to reduce or eliminate a sign or symptom of the disorder, tostabilize the disorder, and/or to reduce or slow further progression ofthe disorder. The subject may be a human subject, or a non-human mammalin some cases.

In one set of embodiments, a composition as described herein is appliedtopically to the skin, e.g., to treat or prevent a disease or conditioncharacterized by a learning or memory disorder, and/or to enhancelearning or memory. For example, the composition may be applied to thecarotid arteries, e.g., for delivery into the head or the brain.Examples of learning and memory disorders include, but are not limitedto, agnosia, Alzheimer's disease, amnesia, mild cognitive impairment(MCI), traumatic brain injury, dementia, Huntington's Disease,Parkinson's Disease, or Wernicke-Korsakoff's Syndrome. In addition, insome cases, the condition may be mentally retardation. In yet anotherset of embodiments, the subject may be normal (or not indicated ashaving a learning or memory disorder), but wish to enhance his or herlearning or memory abilities.

In another set of embodiments, a variety of inflammatory diseases, suchas inflammatory dermatoses may be treated with a composition asdescribed. Inflammatory dermatoses are generally characterized byinflammation in the epidermis and/or dermis. Examples of inflammatorydermatoses include, but are not limited to, psoriasis, atopicdermatitis, eczematous dermatitis, dermatitis, eczema, contactdermatitis, erythema multiforme, pruritus (urticaria), and the like.Some inflammatory dermatoses are autoimmune in nature. In many cases,lesions or eruptions may appear on the skin, and may be acute (lastingdays to weeks) or chronic (lasting months to years). Acute lesions arerelatively common and exhibit a wide range of clinical conditions.Usually, these conditions are triggered by local or systemic immunologicfactors (e.g., allergic reaction).

In addition, various non-limiting examples of systems and methods oftreatment of a disease or condition using a formulation comprisingnitric oxide are disclosed in the following U.S. patent applications,each filed Mar. 13, 2013, and, each incorporated by reference in itsentirety: “Systems and Methods for Delivery of Peptides” (U.S. patentapplication Ser. No. 13/801,402) “Treatment of Skin, Including AgingSkin, to Improve Appearance” (U.S. patent application Ser. No.13/801,446); “Hair Treatment Systems and Methods Using Peptides andOther Compositions” (U.S. patent application Ser. No. 13/801,488); “SkinTanning Using Peptides and Other Compositions” (U.S. patent applicationSer. No. 13/801,518); “Topical Systems and Methods for Treating SexualDysfunction” (U.S. patent application Ser. No. 13/801,543); “ImmuneModulation Using Peptides and Other Compositions” (U.S. patentapplication Ser. No. 13/800,952); “Cardiovascular Disease Treatment andPrevention” (U.S. patent application Ser. No. 13/801,013); “WoundHealing Using Topical Systems and Methods” (U.S. patent application Ser.No. 13/801,061); “Peptide Systems and Methods for Metabolic Conditions”(U.S. patent application Ser. No. 13/801,110); “Methods and Systems forTreating or Preventing Cancer” (U.S. patent application Ser. No.13/801,188); “Compositions and Methods for Affecting Mood States” (U.S.patent application Ser. No. 13/081,240); “Improvement of Memory orLearning Using Peptide and Other Compositions” (U.S. patent applicationSer. No. 13/801,298); and “Brain and Neural Treatments ComprisingPeptides and Other Compositions” (U.S. patent application Ser. No.13/801,345).

In certain aspects of the invention, a composition such as thosedescribed herein can be administered to a subject, such as a humansubject, by rubbing it on the skin of the subject. Without wishing to bebound by any theory, it is believed that phosphatidylcholine provides orfacilitates delivery of ZW1, etc. to the skin, and/or to tissues belowthe skin, allowing the ZW1 to be delivered. In some embodiments, thecomposition can be applied, by rubbing the composition topically againstthe skin, which allows the composition to be absorbed by the skin. Thecomposition can be applied once, or more than once. For example, thecomposition may be administered at predetermined intervals. In someembodiments, for instance, the composition may be applied once per day,twice per day, 3 times per day, 4 times per day, once every other day,once every three days, once every four days, etc. The amount of ZW1necessary to bring about the therapeutic treatment is not fixed per se,and may depend upon factors such as the desired outcome, the type andseverity the disease or condition, the form of SW1, the concentration ofSW1 and/or related compounds present within the composition, etc.

Thus, some embodiments of the invention provide methods of administeringany composition such as discussed herein to a subject. Whenadministered, the compositions of the invention are applied in atherapeutically effective, pharmaceutically acceptable amount as apharmaceutically acceptable formulation. Any of the compositions of thepresent invention may be administered to the subject in atherapeutically effective dose. When administered to a subject,effective amounts will depend on the particular condition being treatedand the desired outcome. A therapeutically effective dose may bedetermined by those of ordinary skill in the art, for instance,employing factors such as those described herein and using no more thanroutine experimentation.

In certain embodiments of the invention, the administration of variouscompositions of the invention may be designed so as to result insequential exposures to the composition over a certain time period, forexample, hours, days, weeks, months, or years. This may be accomplished,for example, by repeated administrations of a composition of theinvention by one or more of the methods described herein, or by asustained or controlled release delivery system in which the compositionis delivered over a prolonged period without repeated administrations.Administration of the composition using such a delivery system may be,for example, by a transdermal patch. Maintaining a substantiallyconstant concentration of the composition may be preferred in somecases.

For certain chronic treatments or therapies, it is contemplated that acomposition as discussed herein may be used to deliver ZW1 to the skinat a relatively high concentration during an initial treatment, and theamount of ZW1 may be lowered or “titrated” down to a relatively lowerconcentration maintenance dose or amount.

In one set of embodiments, compositions described herein can beadministered to a subject in a dosage range from between about 0.01 toabout 10,000 mg/kg body weight/day, about 0.01 to about 5000 mg/kg bodyweight/day, about 0.01 to about 3000 mg/kg body weight/day, about 0.01to about 1000 mg/kg body weight/day, about 0.01 to about 500 mg/kg bodyweight/day, about 0.01 to about 300 mg/kg body weight/day, about 0.01 toabout 100 mg/kg body weight/day.

In one set of embodiments, the dosage may be between about 0.01 mg andabout 500 g, between about 0.01 mg and about 300 g, between about 0.01mg and about 100 g, between about 0.01 mg and about 30 g, between about0.01 mg and about 10 g, between about 0.01 mg and about 3 g, betweenabout 0.01 mg and about 1 g, between about 0.01 mg and about 300 mg,between about 0.01 mg and about 100 mg, between about 0.01 mg and about30 mg, between about 0.01 mg and about 10 mg, between about 0.01 mg andabout 3 mg, between about 0.01 mg and about 1 mg, between about 0.01 mgand about 0.3 mg, or between about 0.01 mg and about 0.1 mg.

In another set of embodiments, the dosage may be at least about 0.01 mg,at least about 0.02 mg, at least about 0.03 mg, at least about mg, atleast about 0.05 mg, at least about 0.1 mg, at least about 0.2 mg, atleast about 0.3 mg, at least about 0.5 mg, at least about 1 mg, at leastabout 2 mg, at least about 3 mg, at least about 5 mg, at least about 10mg, at least about 20 mg, at least about 30 mg, at least about 50 mg, atleast about 100 mg, at least about 200 mg, at least about 300 mg, atleast about 500 mg, at least about 1 g, at least about 2 g, at leastabout 3 g, at least about 5 g, at least about 10 g, etc. In some cases,the dosage may be no more than about 10 g, no more than about 5 g, nomore than about 3 g, no more than about 2 g, no more than about 1 g, nomore than about 500 mg, no more than about 300 mg, no more than about200 mg, no more than about 100 mg, no more than about 50 mg, no morethan about 30 mg, no more than about 20 mg, no more than about 10 mg, nomore than about 5 mg, no more than about 3 mg, no more than about 2 mg,no more than about 1 mg, no more than about 0.5 mg, no more than about0.3 mg, no more than about 0.2 mg, no more than about 0.1 mg, no morethan about 0.05 mg, no more than about 0.03 mg, no more than about 0.02mg, no more than about 0.01 mg, etc. In some cases, combinations of anyof these are also possible, e.g., between about 0.01 mg and about 0.1mg.

The compositions described herein can be used in combination therapywith one or more additional therapeutic agents. For combinationtreatment with more than one active agent, where the active agents arein separate dosage formulations, the active agents may be administeredseparately or in conjunction. In addition, the administration of oneelement may be prior to, concurrent to, or subsequent to theadministration of the other agent. In certain embodiments, theadditional therapeutic agent is present in a provided composition inaddition to ZW1. In other embodiments, the additional therapeutic agentis administered separately from the composition comprising ZW1.

When co-administered with other agents, an “effective amount” of thesecond agent will depend on the type of drug used. Suitable dosages areknown for approved agents and can be adjusted by the skilled artisanaccording to the condition of the subject, the type of condition(s)being treated and the amount of a compound described herein being used.In cases where no amount is expressly noted, an effective amount shouldbe assumed.

In certain embodiments, a composition comprising ZW1 and the additionaltherapeutic agent are each administered in an effective amount (i.e.,each in an amount which would be therapeutically effective ifadministered alone). In other embodiments, a composition comprising ZW1and the additional therapeutic agent are each administered in an amountwhich alone does not provide a therapeutic effect (a sub-therapeuticdose). In yet other embodiments, a composition comprising ZW1 can beadministered in an effective amount, while the additional therapeuticagent is administered in a sub-therapeutic dose. In still otherembodiments, a composition comprising ZW1 can be administered in asub-therapeutic dose, while the additional therapeutic agent isadministered in an effective amount.

As used herein, the terms “in combination” or “co-administration” can beused interchangeably to refer to the use of more than one therapy (e.g.,one or more prophylactic and/or therapeutic agents). The use of theterms does not restrict the order in which therapies (e.g., prophylacticand/or therapeutic agents) are administered to a subject.

Co-administration encompasses administration of the first and secondamounts of the compounds in an essentially simultaneous manner, such asin a single pharmaceutical composition, for example, capsule or tablethaving a fixed ratio of first and second amounts, or in multiple,separate capsules or tablets for each. In addition, suchco-administration also encompasses use of each compound in a sequentialmanner in either order. When co-administration involves the separateadministration of the first amount of a composition as described herein,and a second amount of an additional therapeutic agent, the compoundsare administered sufficiently close in time to have the desiredtherapeutic effect. For example, the period of time between eachadministration which can result in the desired therapeutic effect, canrange from minutes to hours and can be determined taking into accountthe properties of each compound. For example, a composition as describedherein, and the second therapeutic agent can be administered in anyorder within about 24 hours of each other, within about 16 hours of eachother, within about 8 hours of each other, within about 4 hours of eachother, within about 1 hour of each other or within about 30 minutes ofeach other.

More specifically, a first therapy (e.g., a prophylactic or therapeuticagent such as a composition described herein) can be administered priorto (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or12 weeks before), concomitantly with, or subsequent to (e.g., 5 minutes,15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours,12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) theadministration of a second therapy to a subject.

In one set of embodiments, a composition such as is discussed herein maybe applied to the skin of a subject, e.g., at any suitable location. Thecomposition may be contacted using any suitable method. For example, thecomposition may be rubbed on, poured on, applied with an applicator(e.g., a gauze pad, a swab, a bandage, etc.), or the like. In somecases, the composition can be a liquid, a gel, a cream, a lotion, anointment, a solid “stick,” or the like, that can be applied to the skinby hand, for example, by rubbing or spraying.

The compositions of the present invention may additionally comprise oneor more adjunct ingredients, for instance, pharmaceutical drugs or skincare agents. For example, compositions of the invention may includeadjuvants such as salts, buffering agents, diluents, excipients,chelating agents, fillers, drying agents, antioxidants, antimicrobials,preservatives, binding agents, bulking agents, silicas, solubilizers, orstabilizers. Non-limiting examples include species such as calciumcarbonate, sodium carbonate, lactose, kaolin, calcium phosphate, orsodium phosphate; granulating and disintegrating agents such as cornstarch or algenic acid; binding agents such as starch, gelatin oracacia; lubricating agents such as magnesium stearate, stearic acid, ortalc; time-delay materials such as glycerol monostearate or glyceroldistearate; suspending agents such as sodium carboxymethylcellulose,methylcellulose, hydroxypropylmethylcellulose, sodium alginate,polyvinylpyrrolidone; dispersing or wetting agents such as lecithin orother naturally-occurring phosphatides; thickening agents such as cetylalcohol or beeswax; buffering agents such as acetic acid and saltsthereof, citric acid and salts thereof, boric acid and salts thereof, orphosphoric acid and salts thereof; or preservatives such as benzalkoniumchloride, chlorobutanol, parabens, or thimerosal. Suitableconcentrations can be determined by those of ordinary skill in the art,using no more than routine experimentation. Those of ordinary skill inthe art will know of other suitable formulation ingredients, or will beable to ascertain such, using only routine experimentation.

Preparations can include sterile aqueous or nonaqueous solutions,suspensions and emulsions, which can be isotonic with the blood of thesubject in certain embodiments. Examples of nonaqueous solvents arepolypropylene glycol, polyethylene glycol, vegetable oil such as oliveoil, sesame oil, coconut oil, arachis oil, peanut oil, mineral oil,organic esters such as ethyl oleate, or fixed oils including syntheticmono or di-glycerides. Aqueous solvents include water, alcoholic/aqueoussolutions, emulsions or suspensions, including saline and bufferedmedia. Parenteral vehicles include sodium chloride solution,1,3-butandiol, Ringer's dextrose, dextrose and sodium chloride, lactatedRinger's or fixed oils. Intravenous vehicles include fluid and nutrientreplenishers, electrolyte replenishers (such as those based on Ringer'sdextrose), and the like. Preservatives and other additives may also bepresent such as, for example, antimicrobials, antioxidants, chelatingagents and inert gases and the like. Those of skill in the art canreadily determine the various parameters for preparing and formulatingthe compositions of the invention without resort to undueexperimentation.

In some embodiments, a composition such as described herein may beapplied to a surgical device, tool, or other substrate. For example, acomposition of the invention may be applied to sutures, implants,surgical tools, or other substrates that may come into contact withwounded tissue (e.g., cut tissue) during surgery. In some embodiments, acomposition may be provided as a cream or ointment as described in moredetail herein. It also should be appreciated that certain compositionsof the invention may be provided on surgical dressings, bandages, orother material that is to be contacted to a surgical wound.

In one set of embodiments, a composition such as is described herein maybe applied to a material or substrate immediately prior to use on asubject. However, in some embodiments, a material or substrate may beprepared (e.g., packaged, stored, or otherwise prepared) to contain acomposition prior to use. For example, prepackaged bandages or surgicaldevices, sutures, or implants may be prepared and packaged with acoating of a composition such as is described herein. Compositions ofthe invention may be used for human or other animal subjects (male orfemale).

In another aspect, the present invention is directed to a kit includingone or more of the compositions discussed herein. A “kit,” as usedherein, typically defines a package or an assembly including one or moreof the compositions of the invention, and/or other compositionsassociated with the invention, for example, as described herein. Each ofthe compositions of the kit may be provided in liquid form (e.g., insolution), or in solid form (e.g., a dried powder). In certain cases,some of the compositions may be constitutable or otherwise processable(e.g., to an active form), for example, by the addition of a suitablesolvent or other species, which may or may not be provided with the kit.Examples of other compositions or components associated with theinvention include, but are not limited to, solvents, surfactants,diluents, salts, buffers, chelating agents, fillers, antioxidants,binding agents, bulking agents, preservatives, drying agents,antimicrobials, needles, syringes, packaging materials, tubes, bottles,flasks, beakers, dishes, frits, filters, rings, clamps, wraps, patches,containers, and the like, for example, for using, administering,modifying, assembling, storing, packaging, preparing, mixing, diluting,and/or preserving the compositions components for a particular use, forexample, to a sample and/or a subject.

A kit of the invention may, in some cases, include instructions in anyform that are provided in connection with the compositions of theinvention in such a manner that one of ordinary skill in the art wouldrecognize that the instructions are to be associated with thecompositions of the invention. For instance, the instructions mayinclude instructions for the use, modification, mixing, diluting,preserving, administering, assembly, storage, packaging, and/orpreparation of the composition and/or other compositions associated withthe kit. In some cases, the instructions may also include instructionsfor the delivery and/or administration of the compositions, for example,for a particular use, e.g., to a sample and/or a subject. Theinstructions may be provided in any form recognizable by one of ordinaryskill in the art as a suitable vehicle for containing such instructions,for example, written or published, verbal, audible (e.g., telephonic),digital, optical, visual (e.g., videotape, DVD, etc.) or electroniccommunications (including Internet or web-based communications),provided in any manner.

The following documents are incorporated herein by reference: U.S. Pat.No. 8,668,937, issued Mar. 11, 2014, entitled “Topical Nitric OxideSystems and Methods of Use Thereof”; U.S. Pat. No. 8,435,942, issuedApr. 26, 2006, entitled “Methods for Formulating Stabilized InsulinCompositions”; U.S. Pat. No. 7,182,956, issued Feb. 27, 2007, entitled“Stable Topical Drug Delivery Compositions”; U.S. Pat. No. 8,273,711,issued Sep. 25, 2012, entitled “Topical Drug Delivery UsingPhosphatidylcholine”; U.S. patent application Ser. No. 13/801,402, filedMar. 13, 2013, entitled “Systems and Methods for Delivery of Peptides”;U.S. patent application Ser. No. 13/801,446, filed Mar. 13, 2013,entitled “Treatment of Skin, Including Aging Skin, to ImproveAppearance”; U.S. patent application Ser. No. 13/801,488, filed Mar. 13,2013, entitled “Hair Treatment Systems and Methods Using Peptides andOther Compositions”; U.S. patent application Ser. No. 13/801,518, filedMar. 13, 2013, entitled “Skin Tanning Using Peptides and OtherCompositions”; U.S. patent application Ser. No. 13/801,543, filed Mar.13, 2013, entitled “Topical Systems and Methods for Treating SexualDysfunction”; U.S. patent application Ser. No. 13/800,952, filed Mar.13, 2013, entitled “Immune Modulation Using Peptides and OtherCompositions”; U.S. patent application Ser. No. 13/801,013, filed Mar.13, 2013, entitled “Cardiovascular Disease Treatment and Prevention”;U.S. patent application Ser. No. 13/801,061, filed Mar. 13, 2013,entitled “Wound Healing Using Topical Systems and Methods”; U.S. patentapplication Ser. No. 13/801,110, filed Mar. 13, 2013, entitled “PeptideSystems and Methods for Metabolic Conditions”; U.S. patent applicationSer. No. 13/801,188, filed Mar. 13, 2013, entitled “Methods and Systemsfor Treating or Preventing Cancer”; U.S. patent application Ser. No.13/801,240, filed Mar. 13, 2013, entitled “Compositions and Methods forAffecting Mood States”; U.S. patent application Ser. No. 13/801,298,filed Mar. 13, 2013, entitled “Improvement of Memory or Learning UsingPeptide and Other Compositions”; U.S. patent application Ser. No.13/801,345, filed Mar. 13, 2013, entitled “Brain and Neural TreatmentsComprising Peptides and Other Compositions”; U.S. patent applicationSer. No. 13/019,101, filed Feb. 1, 2011, entitled “Method of DeliveringStable Topical Drug Compositions”; U.S. patent application Ser. No.13/926,688, filed Jun. 25, 2013, entitled “Topical Drug Delivery UsingPhosphatidylcholine”; Int. Pat. Apl. Ser. No. PCT/US2014/025822, filedMar. 13, 2014, entitled “Treatment of Skin, Including Aging Skin, toImprove Appearance”; Int. Pat. Apl. Ser. No. PCT/US2014/025913, filedMar. 13, 2014, entitled “Immune Modulation Using Peptides and OtherCompositions”; Int. Pat. Apl. Ser. No. PCT/US2014/025996, filed Mar. 13,2014, entitled “Cardiovascular Disease Treatment and Prevention”; Int.Pat. Apl. Ser. No. PCT/US2014/025572, filed Mar. 13, 2014, entitled“Wound Healing Using Topical Systems and Methods”; Int. Pat. Apl. Ser.No. PCT/US2014/025630, filed Mar. 13, 2014, entitled “Peptide Systemsand Methods for Metabolic Conditions”; Int. Pat. Apl. Ser. No.PCT/US2014/025758, filed Mar. 13, 2014, entitled “Methods and Systemsfor Treating or Preventing Cancer”; Int. Pat. Apl. Ser. No.PCT/US2014/025898, filed Mar. 13, 2014, entitled “Improvement of Memoryor Learning Using Peptide and Other Compositions”; Int. Pat. Apl. Ser.No. PCT/US2014/025820, filed Mar. 13, 2014, entitled “Brain and NeuralTreatments Comprising Peptides and Other Compositions”; and Int. Pat.Apl. Ser. No. PCT/US2014/025705, filed Mar. 13, 2014, entitled “Systemsand Methods for Delivery of Peptides.”

The following examples are intended to illustrate certain embodiments ofthe present invention, but do not exemplify the full scope of theinvention.

Example 1

This example illustrates techniques for preparing compositions inaccordance with one or more embodiments of the invention. Specifically,four formulations suitable as peptide carriers are provided: twofour-phase formulations (HNC 156-43, HNC 156-50), and two single-phaseformulations (HNC 156-47, HNC 159-136).

HNC 156-43

This formulation is formed of: 77.7% water, 6.0% Phospholipon-90G(American Lecithin Company), 0.10% EDTA-Na_(z) (Sigma), 0.1% citricacid, 5.0% isopropyl palmitate (IPP, Kraft Chemicals), 5.0% Promulgen-D(Lubrizol), 3.0% Arlacel-165 (Croda), 1.0% cetearyl alchohol 50/50, 0.5%Dow Corning Fluid 200-10 CST, 0.1% Tocotrienol-50C (Carotech), 0.5%Optiphen Plus (Lotioncrafter), and 1.0% Seppitonic M3 (Seppic).

The individual ingredients are divided between four phases as follows:

Phase 1: water, Phospholipon-90G, EDTA-Na_(z), citric acid.

Phase 2: IPP, Promulgen-D, Arlacel-165, cetearyl alchohol 50/50, DowCorning Fluid 200-10 CST, Tocotrienol-50C.

Phase 3: Optiphen Plus.

Phase 4: Seppitonic M3.

HNC 156-50

This formulation is formed of: 78.3% water, 10.0% Phospholipon-90G(American Lecithin Company), 0.10% EDTA-Na_(z) (Sigma), 0.2% sodiumhyaluronate (1% solution), 3.0% isopropyl palmitate (IPP, KraftChemicals), 3.0% Promulgen-D (Lubrizol), 4.0% Arlacel-165 (Croda), 0.7%cetearyl alchohol 50/50, 0.5% Optiphen Plus (Lotioncrafter), and 0.2%dimethylethanolamine (DMAE, Sigma).

The individual ingredients are divided between four phases as follows:

Phase 1: water, Phospholipon-90G, EDTA-Na_(z), sodium hyaluronate.

Phase 2: IPP, Promulgen-D, Arlacel-165, cetearyl alchohol 50/50.

Phase 3: Optiphen Plus.

Phase 4: DMAE.

The four-phase compositions are generally prepared as follows: Phases 1and 2 are heated to 60° C., added and mixed together, then allowed tocool to 48° C., at which point Phase 3 is added and mixed. The mixturewas then allowed to cool to 38° C., at which point phase 4 is added andmixed.

HNC 156-47

This formulation is formed of: 47% water, 5.0% PEG-200 (Sigma), 45%Phospholipon-90G (American Lecithin Company), 1.0% benzyl alcohol, and2.0% ethyl alcohol.

HNC 159-136

This formulation is formed of: 65% Phospholipon-90G (American LecithinCompany), 18.0% isopropyl palmitate (IPP, Kraft Chemicals), 8% capriccaprylic triglycerides (RITA Corp.), and 9% propanediol (Dupont).

Any of the above compositions are effective as carriers for peptides.For example, the ZW1 peptide may be incorporated into each of the aboveformulations at a concentration of 5.0 mg/cc.

While several embodiments of the present invention have been describedand illustrated herein, those of ordinary skill in the art will readilyenvision a variety of other means and/or structures for performing thefunctions and/or obtaining the results and/or one or more of theadvantages described herein, and each of such variations and/ormodifications is deemed to be within the scope of the present invention.More generally, those skilled in the art will readily appreciate thatall parameters, dimensions, materials, and configurations describedherein are meant to be exemplary and that the actual parameters,dimensions, materials, and/or configurations will depend upon thespecific application or applications for which the teachings of thepresent invention is/are used. Those skilled in the art will recognize,or be able to ascertain using no more than routine experimentation, manyequivalents to the specific embodiments of the invention describedherein. It is, therefore, to be understood that the foregoingembodiments are presented by way of example only and that, within thescope of the appended claims and equivalents thereto, the invention maybe practiced otherwise than as specifically described and claimed. Thepresent invention is directed to each individual feature, system,article, material, kit, and/or method described herein. In addition, anycombination of two or more such features, systems, articles, materials,kits, and/or methods, if such features, systems, articles, materials,kits, and/or methods are not mutually inconsistent, is included withinthe scope of the present invention.

All definitions, as defined and used herein, should be understood tocontrol over dictionary definitions, definitions in documentsincorporated by reference, and/or ordinary meanings of the definedterms.

The indefinite articles “a” and “an,” as used herein in thespecification and in the claims, unless clearly indicated to thecontrary, should be understood to mean “at least one.” The phrase“and/or,” as used herein in the specification and in the claims, shouldbe understood to mean “either or both” of the elements so conjoined,i.e., elements that are conjunctively present in some cases anddisjunctively present in other cases. Multiple elements listed with“and/or” should be construed in the same fashion, i.e., “one or more” ofthe elements so conjoined. Other elements may optionally be presentother than the elements specifically identified by the “and/or” clause,whether related or unrelated to those elements specifically identified.Thus, as a non-limiting example, a reference to “A and/or B”, when usedin conjunction with open-ended language such as “comprising” can refer,in one embodiment, to A only (optionally including elements other thanB); in another embodiment, to B only (optionally including elementsother than A); in yet another embodiment, to both A and B (optionallyincluding other elements); etc.

As used herein in the specification and in the claims, “or” should beunderstood to have the same meaning as “and/or” as defined above. Forexample, when separating items in a list, “or” or “and/or” shall beinterpreted as being inclusive, i.e., the inclusion of at least one, butalso including more than one, of a number or list of elements, and,optionally, additional unlisted items. Only terms clearly indicated tothe contrary, such as “only one of” or “exactly one of,” or, when usedin the claims, “consisting of,” will refer to the inclusion of exactlyone element of a number or list of elements. In general, the term “or”as used herein shall only be interpreted as indicating exclusivealternatives (i.e. “one or the other but not both”) when preceded byterms of exclusivity, such as “either,” “one of,” “only one of,” or“exactly one of.” “Consisting essentially of,” when used in the claims,shall have its ordinary meaning as used in the field of patent law.

As used herein in the specification and in the claims, the phrase “atleast one,” in reference to a list of one or more elements, should beunderstood to mean at least one element selected from any one or more ofthe elements in the list of elements, but not necessarily including atleast one of each and every element specifically listed within the listof elements and not excluding any combinations of elements in the listof elements. This definition also allows that elements may optionally bepresent other than the elements specifically identified within the listof elements to which the phrase “at least one” refers, whether relatedor unrelated to those elements specifically identified. Thus, as anon-limiting example, “at least one of A and B” (or, equivalently, “atleast one of A or B,” or, equivalently “at least one of A and/or B”) canrefer, in one embodiment, to at least one, optionally including morethan one, A, with no B present (and optionally including elements otherthan B); in another embodiment, to at least one, optionally includingmore than one, B, with no A present (and optionally including elementsother than A); in yet another embodiment, to at least one, optionallyincluding more than one, A, and at least one, optionally including morethan one, B (and optionally including other elements); etc.

When the word “about” is used herein in reference to a number, it shouldbe understood that still another embodiment of the invention includesthat number not modified by the presence of the word “about.”

It should also be understood that, unless clearly indicated to thecontrary, in any methods claimed herein that include more than one stepor act, the order of the steps or acts of the method is not necessarilylimited to the order in which the steps or acts of the method arerecited.

In the claims, as well as in the specification above, all transitionalphrases such as “comprising,” “including,” “carrying,” “having,”“containing,” “involving,” “holding,” “composed of,” and the like are tobe understood to be open-ended, i.e., to mean including but not limitedto. Only the transitional phrases “consisting of” and “consistingessentially of” shall be closed or semi-closed transitional phrases,respectively, as set forth in the United States Patent Office Manual ofPatent Examining Procedures, Section 2111.03.

What is claimed is:
 1. An article for transdermal delivery, the article comprising: a composition comprising a carrier and lecithin, the lecithin comprising ZW1 (SEQ ID NO: 1), wherein the composition further comprises no more than about 250 ppm of water by weight of the composition.
 2. The article of claim 1, wherein the ZW1 is present at at least about 0.5% by weight. 3-4. (canceled)
 5. The article of claim 1, wherein the lecithin is present at at least about 0.25% by weight. 6-10. (canceled)
 11. The article of claim 1, wherein the lecithin comprises a phosphatidylcholine. 12-14. (canceled)
 15. The article of claim 11, wherein at least some of the phosphatidylcholine comprises a polyenylphosphatidylcholine. 16-18. (canceled)
 19. The article of claim 1, wherein the lecithin is present as a liquid crystal structure. 20-23. (canceled)
 24. The article of claim 1, wherein composition comprises no more than about 100 ppm of water by weight of the composition. 25-29. (canceled)
 30. The article of claim 1, wherein the composition further comprises molecular nitric oxide. 31-32. (canceled)
 33. The article of claim 1, wherein the composition is a gel.
 34. (canceled)
 35. The article of claim 1, wherein the composition is substantially transparent. 36-37. (canceled)
 38. An article for transdermal delivery, the article comprising: a composition comprising a carrier and lecithin, the lecithin comprising ZW1 (SEQ ID NO: 1), wherein the composition is stable at room temperature.
 39. The article of claim 38, wherein the ZW1 is present at at least about 0.5% by weight. 40-41. (canceled)
 42. The article of claim 38, wherein the lecithin is present at at least about 0.25% by weight. 43-44. (canceled)
 45. The article of claim 38, wherein the lecithin comprises a phosphatidylcholine. 46-48. (canceled)
 49. The article of claim 45, wherein at least some of the phosphatidylcholine comprises a polyenylphosphatidylcholine.
 50. The article of claim 38, wherein the lecithin is present as a liquid crystal structure. 51-54. (canceled)
 55. The article of claim 38, wherein composition comprises no more than about 250 ppm of water by weight of the composition.
 56. (canceled)
 57. The article of claim 38, wherein the composition further comprises molecular nitric oxide. 58-59. (canceled)
 60. The article of claim 38, wherein the composition is a gel.
 61. (canceled)
 62. The article of claim 38, wherein the composition is substantially transparent. 63-97. (canceled) 